The Emergence of Universal Immune Receptors for Highly Personalized T cell Therapy

Transcription

1 The Emergence of Universal Immune Receptors for Highly Personalized T cell Therapy INNOVATIVE APPROACHES TO T CELL BASED THERAPIES Daniel Powell Ph.D. Dept of Pathology and Laboratory Medicine Abramson Cancer Center Perelman School of Medicine University of Pennsylvania

2 Chimeric Antigen Receptor Approach A. B. Michael H. Kershaw,et al. Nat Rev Immunol.2005 Two-signal model of T-cell activation. C. 1 st 2 nd 3 rd GENERATION CD3z CD28/ 4-1BB CD28 Advantages of CAR T cells: 1. De novo generation of tumor-reactive T cells 2. MHC non-restricted 3. Surface antigen detection 4. Combines signal Potential universal use CD3z 4-1BB CD3z

3 CD19 CAR T cell-mediated tumor regression UPENN UPENN SB, NCI MSKCC SB, NCI

4 A strategy for personalized immunotherapy of cancer using gene-modified T cells Challenges: Cost of gene transfer vectors Potential for toxicity against normal tissue bearing low level antigen Long lasting persistence Potential immunogenicity of receptors of non-human origin Outgrowth of antigen loss variant cancers Rosenberg SA, 2011, Sci Transl Med. 4;127: sp8.

5 Fixed antigen specificity in conventional CAR design One CAR: One Antigen Specificity Tumors are highly heterogeneous in the expression of cell surface antigens CAR therapy remains susceptible to tumor evasion (e.g. loss of antigen expression) 5

6 Creation of a Universal Immune Receptor platform for targeting unlimited antigen specificities A STANDARDIZED ADAPTABLE IMMUNE RECEPTOR PLATFORM unlimited potential antigen specificity for patient-tailored T cell therapy flexibility in targeting targeting sequential and/or simultaneous to address antigen loss variants incorporate accessory signaling (e.g. costimulation) ex vivo selection of specific T cell subsets to be used (e.g. CD8, CD4, Th17, Treg, etc.) allow for T cell redirection via a wide array of antigen-specific molecules (e.g. scfvs, antibodies, aptamers, receptors, ligands, small molecules)

7 Principle of Biotin-Binding Immune Receptor (BBIR) Platform T Cell T cell signaling domains Avidin domains BBIR T Cell Antibody and/or scfv Biotin TAA BBIR TAA Painted TAAs Tumor Biotinylated scfvs or antibodies enable immuno-recognition by a Biotin-Binding Immune Receptor (BBIR) K. Urbanska and D.J. Powell Jr. Oncoimmunology 2012

8 Construction of Biotin-Binding Immune Receptors - BBIRs dcav BBIR-z dcav BBIR-28z dcav; biotin binding domain cav cav cav cav CD3z CD28 mcav; biotin binding domain CD3z GFP 2A mcav BBIR-z GFP 2A mcav BBIR-28z cav cav CD3z CD28 CD3z pelns based Lentiviral vectors leader hinge TM Primary human T cells efficiently transduced to express mcav BBIR (monomeric) or dcav BBIR (dimer) on their cell surface dcav-bbir z dcav-bbir 28z GFP 2A mcav-bbir z GFP 2A mcav-bbir 28z GFP 76 % 66 % 69% 75 % 92 % cav Ab-FITC mcav-gfp GFP

9 IFNg pg/ml IFNg pg/ml BBIR+ T cells recognize biotinylated antibodies and/or bio-bodies in vitro GFP BBIR-z 5000 mcav BBIR-z dcav-bbir-z BBIR-28z GFP Ab Conc. 10ng/well h-mesothelin (10ng/well) Immunorecognition of multiple biotinylated molecules Immunorecognition of painted antigens P4 Biobody Bio-IgG Ab. P4 Biobody Bio-K1 Ab. Meso Meso Meso Immobilized antibodies Immobilized antigen (10ng/well) Urbanska K. et al. Cancer Research 2012

10 IFNg pg/ml Soluble Biotin does not activate or inhibit BBIR + T cells function Biotin Conc. 0nM 3nM 40nM empty Bio-IgG1 IgG1 P4 Biobody P4scFv labeled rh-mesothelin protein BBIR T cells show functionality in the presence of free Biotin at the physiological levels Single bolus injection of dcav has been shown to be safe and non-immunogenic

11 IFNg pg/ml IFNg pg/ml BBIR T cells recognize painted antigens on cancer cell surface A1847 (EpCAM+) ng 100 ng 10 ng A1847 (EpCAM+) BBIR-28z GFP ng ng Isotype 5000 Surface Bio-EpCAM Ab A1847 labeled with Bio-EpCAM (ng) R² = BBIR-28z GFP Direct correlation between intensity of single antigen labeling and immune recognition by BBIR + T cells R² = Specific MFI (*10 3 ) Urbanska K. et al. Cancer Research 2012

12 Universality: Retargeting BBIR T cells using biotinylated Ab or scfvs against multipletaas Mesothelin Fra EpCAM Bio-K1 Ab Bio-P4 scfv Bio-MOV18 Ab Bio-EpCAM Ab AE17 / mesothelin AE17 / Fra A1847/ EpCAM + /Fra + /mesothelin +

13 IFNg pg/ml IFNg pg/ml BBIR+ T cells can be redirected against multiple painted antigens expressed on cancer cell surface BBIR-28z AE17/ Mesothelin + AE17/ FRa + BBIR-28Z GFP A1847/FRa + Mesothelin + EpCAM + Th1 type cytokine production by BBIR-28z In response to painted A1847 (FRa + Mesothelin + EpCAM + ) A1847 cells

14 Cell count Flexibility of BBIR allows for sequential targeting of distinct cancer antigens TAAs EpCAM FRa BBIR T cells re-directed against 1 st target EpCAM via Bio-EpCAM Ab AE17 / Fra + A1847/ EpCAM + /Fra + BBIR T cells re-directed against 2 nd target FRa via Bio-MOV18 Ab Tumor elimination Abs to TAAs targeted by BBIR-28z Urbanska K. et al. Cancer Research 2012

15 BBIR BBIR BBIR+ T cells kill human ovarian cancer cells via EpCAM and/or Mesothelin redirection in vitro Specific killing of A1847 (FRa + Mesothelin + EpCAM + ) cells by EpCAM redirected BBIRs Specific killing of A1847( FRa + Mesothelin + EpCAM + ) cells by Mesothelin redirected BBIRs E:T ratio E:T ratio Universality of BBIR platform allows for a specific BBIR + T cells mediated tumor lysis via painting variable antigens expressed on the cancer cell surface Urbanska K. et al. Cancer Research 2012

16 Tumor Volume (mm3) Antitumor activity of redirected BBIR + T cells BBIR Bio-EpCAM BBIR Bio-IgG1 * (IT) Administration of T cells + Bio-EpCAM Ab * * * Days post tumor injection Tumors were significantly smaller in mice receiving BBIR + T cells and Bio-EpCAM Ab. Urbanska K. et al. Cancer Research 2012

17 Summary The BBIR platform represents a universal immune receptor approach for the targeting of gene-modified T cells to diverse and multiple antigens via interaction with antigen-bound biotinylated molecules, either simultaneously or sequentially. Screening platform Sequential targeting Targeting multiple TAAs Biotinylated: scfv Ab Ligands Aptamers Targeting 1 st TAA Redirection of BBIR via Abs of distinct antigen specificity Immunoediting Antigen escape Targeting 2 nd TAA CAR construction TAA selection for immunotherapy Antitumor response Secondary infusion of Abs of distinct antigen specificity Antitumor response Other platforms remain in development (higher affinity binding; fully human construsts; novel binding partners) Antitumor response

18 CONCLUSION Adoptive immunotherapy has emerged as a promising approach for the treatment of advanced cancer. Gene-modified T cells represent an offthe-shelf approach to effective treatment of a wide variety of cancers. Next generation strategies to T cell based therapy will be required to improve efficacy, safety and broaden patient access to this promising approach.

19 ACKNOWLEDGEMENTS Powell Lab: Kasia Urbanska Evripidis Lanitis Mathilde Poussin Degang Song Rachel Lynn Jenessa Smith Qunrui Ye Andrew Best Caitlin Stashwick Keith Schutzky Thomas Garrabrant Key Collaborators: Nathalie Scholler Carl June Gwenn Danet UPENN OCRC & TRP ACC members Clinical Staff Support: NIH (R21CA152540) NIH (R01CA168900) SPORE (P50 CA083638) PPG Ovarian Cancer Research Fund Basser Research Center for BRCA Research Marsha Rivkin Foundation Sandy Rollman Foundation